J Otolaryngol Ophthalmol Shandong Univ ›› 2018, Vol. 32 ›› Issue (4): 53-57.doi: 10.6040/j.issn.1673-3770.0.2017.301

Previous Articles     Next Articles

Up-regulation of miR-200b may inhibit epithelial mesenchymal transition process to prevent the proliferation, migration, and invasion of human laryngeal cancer Hep-2 cells

LI Guobin, ZHANG Zhancheng, WANG Xinyan   

  1. Department of Otolaryngology, Fourth Peoples Hospital of Jinan, Jinan 250031, Shandong, China
  • Received:2017-07-11 Online:2018-07-20 Published:2018-07-20

Abstract: Objective We investigated the effects of up-regulation of miR-200b on the proliferation, migration, and invasion of human laryngeal cancer Hep-2 cells. Methods Human laryngeal carcinoma Hep-2 cells were cultured and randomly divided into the miR-200b mimic group, miR-control sequence group, and blank control group. The expression of miR-200b was detected by real-time fluorescence quantitative PCR. The MTT assay was used to detect cell proliferation. The Transwell method was used to detect cell migration and invasion. The expression of E-cadherin, N-cadherin, and β-catenin proteins was detected by western blotting. Results Compared to the blank control group and miR-control sequence group, the relative expression level of miR-200b in the miR-200b mimic group was significantly increased(F=70.766, P<0.001). Compared to the blank control group [(0.22±0.05),(0.45±0.07),(0.59±0.06),(0.72±0.08), and(0.85±0.07)] and miR-control sequence group [(0.24±0.06),(0.46±0.08),(0.61±0.07),(0.70±0.05), and(0.84±0.06)], the absorbance A values at 24, 48, 72, and 96 h in the miR-200b mimic group [(0.21±0.04),(0.29±0.06),(0.40±0.04),(0.53±0.07), and(0.58±0.05)] were decreased and the differences were statistically significant(F=0.134, 6.449, 37.299, 5.352, and 29.921, P=0.876, 0.010, 0.000, 0.018, and 0.000, P<0.05). Compared to the blank control group [(140.2±2.3),(127.9±6.0)] and miR-control sequence group [(141.0±1.2),(130.4±7.4)], the number of migrating cells and number of invasive cells in the miR-200b mimic group [(98.5±2.4),(90.9±2.8)] were decreased and the differences were statistically significant(F=845.523, 88.859, P both <0.001). Compared to the blank control group [(0.35±0.07),(0.54±0.10),(0.76±0.12)] and miR-control sequence group [(0.24±0.03),(0.60±0.14),(0.65±0.24)], the relative expression level of E-cadherin protein in miR-200b mimic group cells(0.54±0.14)was increased, whereas the relative expression levels of N-cadherin and β-catenin proteins [(0.31±0.10),(0.35±0.06)] were decreased; these differences were statistically significant(F=17.287, 10.083, 10.434, P<0.001, 0.002, 0.001). Conclusion Up-regulation of miR-200b gene expression in laryngeal squamous cell carcinoma may reduce cell proliferation and inhibit cell migration and invasion. The mechanism may be related to inhibition of the epithelial mesenchymal transition process.

Key words: miR-200b, Epithelial-mesenchymal transition, Cell proliferation, Cell invasion, Laryngeal carcinoma

CLC Number: 

  • R739.6
[1] Gorphe P, Tao Y, Blanchard P, et al. Relationship between the time to locoregional recurrence and survival in laryngeal squamous-cell carcinoma[J]. Eur Arch Otorhinolaryngol, 2017, 274(5):2267-2271.
[2] Shi X, Hu WP, Ji QH. Development of comprehensive nomograms for evaluating overall and cancer-specific survival of laryngeal squamous cell carcinoma patients treated with neck dissection[J]. Oncotarget, 2017, 8(18):29722-29740.
[3] Sliwinska A, Kasinska MA, Drzewoski J. MicroRNAs and metabolic disorders-where are we heading?[J]. Arch Med Sci, 2017, 13(4):885-896.
[4] Humphries B, Yang C. The microRNA-200 family: small molecules with novel roles in cancer development, progression and therapy[J]. Oncotarget, 2015, 6(9):6472-6498.
[5] Yao CX, Wei QX, Zhang Y, et al. miR-200b targets GATA-4 during cell growth and differentiation[J]. RNA Biol, 2013, 10(4):465-480.
[6] Iser IC, Pereira MB, Lenz G, et al. The epithelial-to-mesenchymal transition-like process in glioblastoma: an updated systematic review and in silico investigation[J]. Med Res Rev, 2017, 37(2):271-313.
[7] Wang M, Wu CP, Pan JY, et al. Cancer-associated fibroblasts in a human HEp-2 established laryngeal xenografted tumor are not derived from cancer cells through epithelial-mesenchymal transition, phenotypically activated but karyotypically normal[J]. PLoS One, 2015, 10(2):e0117405.
[8] Yao CX, Wei QX, Zhang YY, et al. miR-200b targets GATA-4 during cell growth and differentiation[J]. RNA Biol, 2013, 10(4):465-480.
[9] Fodor M, Cosgarea M, Fodor L. Simultaneous total esophagectomy and anterior mediastinal tracheostomy for recurrent laryngeal cancer extended to the superior mediastinum[J]. Chirurgia(Bucur), 2017, 112(1):63-67.
[10] 席盼盼. 凋亡相关基因及其与喉癌关系的研究进展[J]. 蚌埠医学院学报, 2016, 41(1):137-140. XI Panpan. Advances in the study of apoptosis-related genes and their relationship with laryngeal[J]. J Bengbu Med Coll, 2016, 41(1):137-140.
[11] Yu X, Li Zh. The role of MicroRNAs expression in laryngeal cancer[J]. Oncotarget, 2015, 6(27):23297-23305.
[12] 唐峰波, 杨铭. MiR-200家族与消化道肿瘤的相关研究[J]. 中华临床医师杂志(电子版), 2016, 10(1):110-115. TANG Fengbo, YANG Ming. Allied study between miR-200 family and digestive system neoplasms[J]. Chin J Clin(Electr Edit), 2016, 10(1):110-115.
[13] Pendlebury A, Hannan NJ, Binder N, et al. The circulating microRNA-200 family in whole blood are potential biomarkers for high-grade serous epithelial ovarian cancer[J]. Biomed Rep, 2017, 6(3):319-322.
[14] 罗卫民, 罗湘玉, 郭家龙, 等. miR-200b靶向DNMT3A抑制非小细胞肺癌细胞增殖与诱导凋亡[J]. 天津医药, 2016, 44(8):984-988. LUO Weimin, LUO Xiangyu, GUO Jialong, et al. miR-200b suppresses proliferation and induces apoptosis in non-small cell lung cancer cells by targeting DNMT3A[J]. Tianjin Med J, 2016, 44(8):984-988.
[15] Wang N, Wei L, Huang Y, et al. miR520c blocks EMT progression of human breast cancer cells by repressing STAT3[J]. Oncol Rep, 2017, 37(3):1537-1544.
[16] Asakura T, Yamaguchi N, Ohkawa K, et al. Proteasome inhibitor-resistant cells cause EMT-induction via suppression of E-cadherin by miR-200 and ZEB1[J]. Int J Oncol, 2015, 46(5):2251-2260.
[17] 李艳霞, 廖艳, 丁峰, 等. Notch-1和微小RNA-200家族在胰腺癌上皮-间充质转化中的作用[J]. 中华实验外科杂志, 2016, 33(7):1792-1796. LI Yanxia, LIAO Yan, DING Feng, et al. The mechaniSills of Notch-1 and microRNA-200 family in epithelial-mesenchymal transition of pancreatic cancer[J]. Chin J Exp Surg, 2016, 33(7):1792-1796.
[18] Rogers CD, Saxena A, Bronner ME. Sip1 mediates an E-cadherin-to-N-cadherin switch during cranial neural crest EMT[J]. J Cell Biol, 2013, 203(5):835-847.
[19] 宋盼盼, 钱晓云, 周涵, 等. 上皮钙黏蛋白和神经钙黏蛋白以及β-连环蛋白在喉鳞状细胞癌中的表达及临床意义[J]. 中华耳鼻咽喉头颈外科杂志, 2016, 51(6):440-445. SONG Panpan, QIAN Xiaoyun, ZHOU Han, et al. Expression of E-cadherin, N-cadherin, β-catenin and their clinical significance in laryngeal carcinoma[J]. Chin J Otorhinolaryngol Head Neck Surg, 2016, 51(6):440-445.
[20] Liv ZD, Wang HB, Liu XP, et al. Silencing of Prrx2 inhibits the invasion and metastasis of breast cancer both in Vitro and in Vivo by reversing epithelial-mesenchymal transition[J]. Cell Physiol Biochem, 2017, 42(5):1847-1856.
[1] YE Shufeng, ZHANG Xu, SU Kai, LI Hejie, TENG Bo. Laryngocelesecondary to Laryngectomy:a case report and literature review [J]. J Otolaryngol Ophthalmol Shandong Univ, 2018, 32(5): 117-118.
[2] WANG Hui, WEN Yan, YAN Li, YU Xiaoming, NING Hong. The effect of transforming growth factor-β1 on epithelial-mesenchymal transition in human lens epithelial cells. [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2017, 31(4): 76-79.
[3] XU Yuanteng, CHEN Ruiqing, LIN Gongbiao, FANG Xiuling, YU Shujuan, LIANG Xiaohua, ZHANG Rong. Effect of silencing PDCD4 gene on proliferation of Hep-2 cells and the expression of β-catenin by RNA interference technique. [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2016, 30(5): 110-114.
[4] LI Zewen, GUO Junyu, ZHOU Jie, YAN Fubo, YANG Zhimin, DING Zhuhua. Relationship between gastroesophageal reflux disease and laryngeal carcinoma: a meta-analysis. [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2016, 30(3): 40-46.
[5] LI Fu, ZHAO Shuyou. Expression of tumor suppressor in lung cancer 1 in laryngeal squamous cell carcinoma and its significance [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2015, 29(6): 36-38.
[6] CHEN Xiaoling, FEI Bing, LI Xianbin, WU Hao. Construction and identification of small interfering RNA targeting silence Trop2 expression in human laryngeal carcinoma cell line HEP2 [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2015, 29(3): 51-53.
[7] GUO Zhi-juan1, LI Pei-hua1, ZHANG Xiao-wen2. Prognostic significance of epithelial-mesenchymal transition state in patients with chronic sinusitis treated under endoscopy [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2014, 28(1): 34-38.
[8] PAN Chun-chen1, ZHANG Xiao-peng2, LAI Hai-chun3, WANG De-sheng3. Expression and  relationship of HMGA1 and Ezrin in laryngeal carcinoma [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2012, 26(2): 27-31.
[9] ZOU Ji-dong, XU Wei, L Zheng-hua, CAO Hong-yuan. Correlation between maspin and p53 proteins in laryngeal carcinoma [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2011, 25(6): 16-18.
[10] LI Da-wei, DONG Pin, GAO Shang. Significance of apoptotic and proliferative indices and  P-glycoprotein expression in laryngeal squamous cell carcinoma [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2011, 25(4): 8-12.
[11] Qian Xiaoyun1, Zhang Yifen2, Yu Chenjie1, Ding Xiaoqiong1, Qin Yang1, Qin Xiaoming1, Gao Xia1. Detection of micrometastasis in cervical lymph nodes from laryngeal or laryngopharyngeal carcinoma and preliminary research on the clinical significance of micrometastasis [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2010, 24(3): 9-14.
[12] GAO Chun-rong,YAN Yong-hong,CHAI Xiang-jun . Pathogens in sputum in laryngeal cancer patients after tracheotomy [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2007, 21(6): 498-499 .
[13] SUN Xin,JI Wen-yue . Long-term curative effect of the KTP laser for laryngeal carcin [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2007, 21(5): 393-395 .
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!